Preparation method for irritation reducing fermentation tobacco extract, and applications of irritation reducing fermentation tobacco extract in recombinant tobacco
A tobacco leaf extract and tobacco leaf technology are applied in the field of reconstituted tobacco leaf additives, which can solve the problems of high irritation, unpleasant taste, heavy miscellaneous gas, etc., and achieve the effects of improving the irritation, reducing the irritation of smoke, and filling the smoke.
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Embodiment 1
[0025] The preparation method of the microbial plant Raoultella planticola VP4-4 CCTCC M 2012005 biotransformation production fermented tobacco leaf extract of the present invention is as follows:
[0026] 1) Crush the tobacco leaves to 100 mesh, then reflux extract the tobacco powder and water at a mass ratio of 1:60 for 3 hours, then filter, repeat the reflux once to extract the filtrate, and combine the filtrates to obtain the tobacco leaf extract, which is then sterilized;
[0027] 2) Strain activation: inoculate Raoultella planticola VP4-4 CCTCC M 2012005 on the slant medium, and culture at 38°C for 24 hours to obtain the activated strain, and place it in a refrigerator at 4°C;
[0028] The slant culture medium is: wort juice: 120mL, sucrose: 20g / L, agar: 15g / L.
[0029] 3) Preparation of seed culture solution: inoculate the activated strains obtained in step 2) into the seed culture medium, culture at 25°C with shaking at 100r / min for 72 hours, and transfer continuously ...
Embodiment 2
[0034] The preparation method of the microbial plant Raoultella planticola VP4-4 CCTCC M 2012005 biotransformation production fermented tobacco leaf extract of the present invention is as follows:
[0035] 1) Crush the tobacco leaves to 100 mesh, then reflux extract the tobacco powder and water at a mass ratio of 1:40 for 3 hours, then filter, repeat the reflux for 3 times to extract the filtrate, and combine the filtrates to obtain the tobacco leaf extract, and sterilize it;
[0036] 2) Strain activation: inoculate Raoultella planticola VP4-4 CCTCC M 2012005 on the slant medium, and culture at 30°C for 72 hours to obtain the activated strain, and place it in a refrigerator at 4°C;
[0037] The slant culture medium is: wort juice: 120mL, sucrose: 20g / L, agar: 15g / L.
[0038]3) Preparation of seed culture solution: Inoculate the activated strains obtained in step 2) into the seed culture medium, culture at 32°C with shaking at 100r / min for 48 hours, and transfer continuously fo...
Embodiment 3
[0043] The preparation method of the microbial plant Raoultella planticola VP4-4 CCTCC M 2012005 biotransformation production fermented tobacco leaf extract of the present invention is as follows:
[0044] 1) Crush the tobacco leaves to 50 mesh, then reflux extract the tobacco powder and water at a mass ratio of 1:20 for 1 hour, then filter, repeat the reflux once to extract the filtrate, and combine the filtrates to obtain the tobacco leaf extract, which is then sterilized;
[0045] 2) Strain activation: inoculate Raoultella planticola VP4-4 CCTCC M 2012005 on the slant medium, and culture at 25°C for 24 hours to obtain the activated strain, and place it in the refrigerator;
[0046] The slant culture medium is: wort juice: 120mL, sucrose: 20g / L, agar: 15g / L.
[0047] 3) Preparation of seed culture solution: Inoculate the activated strain obtained in step 2) into the seed medium, shake and culture at 100r / min for 72 hours at 25°C, and transfer once continuously to obtain the ...
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